: Project 3 will consist of three sub-projects, which could lead to significant improvements for a therapeutic vaccine strategy. First, both gag and gpl60 genes will be engineered so that they are expressed from the same replicon RNA. Given the high expression levels of the VEE replicons and results of other studies, we anticipate that gag-env particles will bud from the plasma membranes of cells infected with such VRPs. Particulate antigens are often superior immunogens. As these HIV virus-like particles should be very similar in structure to immature and mature forms of HIV itself, it is possible that immunization with these VRP could induce CTL as well as higher titer neutralizing antibodies which might be capable of neutralizing primary isolates. The second therapeutic immunization strategy involves using replicon RNA encoding HIV antigens or VRP encoding HIV antigens to transduce a patient?s autologous dendritic cells cultured ex vivo followed by reintroduction of these transduced cells into the patient as a vaccine. The third strategy to be explored is the use of molecular adjuvant proteins covalently linked to an HIV immunogen. One method is for accelerated processing of CTL epitopes for presentation using mono-ubiquitinated immunogens. Constructs expressing a ubiquitin-Gag fusion protein will be tested for their ability to induce a stronger cellular immune response compared to Gag alone. In a second method, gpl40 will be linked to Cd3 in an effort to enhance the neutralizing response in the context of the VEE vectors.